Attempts were made to develop a technique to rear the phyllosoma larvae of Panulirus homarus. The biological characters like fecundity, hatching percentage, larval morphological changes, feed inputs and moulting frequency till the fourth moult were studied. Morphometric and meristic characters of the larvae were also studied till the 42ndday. The larval output was directly proportional to the size of the gravid brood stock. Relationship between the duration of culture X and length of the larvae Y were shown by the relationships Y intercept = 0.5780 ± 0.1074 and X intercept = 0.7283 r2 = 0.8519 . There was significant p 0.0001 positive relationship between total length TL and carapace width CW of phyllosoma larvae. S. Lazarus | J. C. Nisha | R. Thangaraja "Studies on the Phyllosoma Larva of the Indian Rock Lobster, Panulirus Homarus Linnaeus, 1758" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-4 | Issue-4 , June 2020, URL: https://www.ijtsrd.com/papers/ijtsrd31676.pdf Paper Url :https://www.ijtsrd.com/biological-science/molecular-biology/31676/studies-on-the-phyllosoma-larva-of-the-indian-rock-lobster-panulirus-homarus-linnaeus-1758/s-lazarus
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The brooders of first two maturationstages,withorangeand
red coloured eggs, were not stocked in the hatching tank.
They were maintained in the maturation tank and fed with
clams and muscles and water exchange till the egg reached
the third stage (i.e.,) the brown coloured egg stage. The 3rd
stage exogenous animal with dark browneggsobtainedboth
from natural source and maturation tanks were transferred
in to the hatching tank for the release of youngonesfrom the
eggs. In the maturation tank, the brooders were fed with
squid and mussel meat at a rate of 20% body weight and
50% water exchange was carriedoutdaily.Thestressduring
this stage was minimized in order to prevent the premature
shedding of eggs.
The hatching was confirmed in the tanks by the vigorous
pleopod movements of the mother lobster and following
larval release in the hatching tank. The hatched larvae were
separated intermittently from the spawning tank and
stocked in larval rearing tanks of 200 L capacityattherateof
10 nos./L stocking density, where the larvae were fed with
Nannochloropsis, an ultra plankton, at the rate of 1 lakh
cells/ml concentration. From the second day onwards, the
larvae were fed with newly hatched Artemia nauplii and
micro algae. The physical parameters maintained duringthe
culture were temperature 30 ± 2° C, salinity 35 ̊/°°, dissolved
oxygen 5 mg/l and pH 8.0 ± 2 (Lazarus et al., 2000).
Following the method of Babu et al. (2001a), cradle aeration
system was improvised for a homogenized and adequate air
supply to the rearing system. About 50-90% of sea water
exchange was carried out daily. The morphological changes
observed in antennules, antenna, head, eye, eyestalk, I
maxilliped, II maxilliped, III maxilliped, I, II, III and IV
pereiopod, setae, spines and cephalothorax were observed
using light microscope and all the morphological variations
were drawn using camera lucida.Thefecundity,larval moult,
growth rate and survival rates were also recorded.
The fecundity was recorded in relation with spawner size.
The exogenous spawners of a visually acceptable size were
allowed to release their eggs as hatched out larvae. After the
release of larvae, both released larvae and unhatched eggs
were counted. From the counted value, the total fecundity
were calculated. Percentage ofsurvival wascalculatedonthe
basis of daily mortality. The growth rate was determined
microscopically by measuring the total length (TL) and
carapace width (CW) of larvae during every moult.
Statistical Analysis:
Coefficient of regression analysis was performed for total
length (CL) and carapace width (CW). Data analysis were
based on mean ± SEM. Turkey’s Multiple Comparison Test
was performed for moulting frequency and fecundity with
95% confidence level. This statistical analysis was
performed by using the software Graph Pad Version 4.0.
RESULTS
Metamorphosis
Before 1st moult
In the newly hatched larvae, the head was cone-shaped. The
antennules and antennae were uniramous. The eyes were
without eyestalk. Mandibles slightly developed. First and
second maxillae were seen as rudiments. Second maxilliped
was uniramous with 4 segments. First pereiopods were
biramous and segmented. Endopod 3 segmented, ends with
long narrow spine, surrounded by a group of 7 small spines.
The terminal segment was spiny and the 2nd pereiopod
rudimentary (Fig. 1).
Afer 1st moult
Head was slightly narrower at the anterior region and
broader at the posterior region. Antennuleuniramous,spiny
and unsegmented with 3 aesthetases and 2 setae in the
terminal region. Antennae uniramous, unsegmented with 2
pairs of setae. Distinct eyestalk was observed. Mandibles
slightly developed. 1st and 2nd maxilla were seen
rudimentary.2nd maxilliped uniramous,foursegmented. Last
segment had 2 setae and third segment had 6 setae. 1st
pereiopod biramous, endopod 3 segmented, ends in a long
narrow spine, endopod 7 segmented with6pairsofplumose
setae and the 4th pereiopod seen as rudiment (Fig.2).
After 2nd moult
Head, narrow in the anterior region and broader at the
posterior region. Antennae and antennules uniramous.
Distinct eye stalk. 1st and 2nd maxilla rudiments, mandibles
slightly developed. 1st maxilliped uniramous, 2 segmented.
Second maxilliped uriramous, 5 segmented and segments
had 3 setae. Third segment had 6 setae. First pereiopod
biramous, exopod 8 segmented with 7 pairs of plumose
setae. Towards the end of the stage, 9 segment and 8 pairs of
plumose setae were seen. Fourth pereiopod was
rudimentary (Fig. 3).
After 3rd moult
Head, narrower in the anterior region and broader at the
posterior region. Antennae and antennules uniramous.
Distinct and elongated eye stalk. First and second maxilla
and mandibles slightly developed. Second maxilliped
uniramous, 5 segmented, last segment had 3 setae and third
segment had 6 setae. First pereiopod biramous, exposed, 10
segmented with 9 pairs of plumose setae. Fourth pereiopod
slightly developed (Fig.4).
After 4th moult
There was no change in the shape of cephalic shield,
antennae and antennules, distinct and elongated eye stalk.
1st and 2nd maxillae and mandibles slightly developed. First
maxillipede 3segmented,secondmaxillipedeuniramousand
5 segmented, last segment had 3 setae. Third maxillipede
bigamous with exopod setae. First and second pereiopods
biramous, 7 segmented with 9 pairs of plumose setae in the
exopod. Endopod ends with long narrow spine surrounded
by a group of small spines. The terminal segment was spiny,
and the third periopod was biramous with five pairs of
exopod setae. Fourth pereiopod slightly developed and bi-
segmented (Fig. 5).
Growth rate
The growth parameter of therelationshipoftotal length(TL)
and carapace width (CW) of phyllosoma larvae is shown in
the Fig. 6. Measurement of total lengthandcarapacewidthof
the larvae was carried out up to 42 days. The newly hatched
larvae had a total length and carapace width of 1.3280 ±
0.0360 mm and 0.6241± 0.0561mm respectively.Onthe11th
day, the larval length and width observed were 1.6144 ±
0.1290 mm and 0.7960 ± 0.063mm respectively. On 21st day
the measured length and width were 1.7004 ± 0.0993mm
and 0.8266 ± 0.0450 mm respectively. On the 31st day the
standard length and widthof thelarvaeobservedwere2.283
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± 0.0116 mm and 1.0346 ± 0.0565 mm respectively and on
the 41st day the standard total length and carapace width of
the larvae observed were 2.894 ± 0.6210 mm and 2.017 ±
0.1432 mm respectively (Fig. 6). The slope of the regression
data showed positive and highly significant (p<0.0001)
relationship between total length (TL) and carapace width
(CW) of phyllosoma larvae.
Fig.1. Newly hatched Phyllosoma larva
Fig.2. Phyllosoma larva after 1st moult
At – Antenna, An – Antennule, Max III – Maxillipede III, P1 –
Pereiopod I, P2 – Pereiopod II, P3 – Pereiopod III, P4 –
Pereiopod IV, Ab – Abdomen, CS – Chephalic shield, ExB –
Exopod budding
Fig.3. Phyllosoma larva after 2nd moult
Fig.4. Phyllosoma larva after 3rd moult
At – Antenna, An – Antennule, Max I – Maxillipede I, Max II –
Maxillipede II, Max III – Maxillipede III,P1– PereiopodI,P2–
Pereiopod II, P3 – Pereiopod III, P4 – Pereiopod IV, Ab –
Abdomen, CS – Chephalic shield
Fig.5. Phyllosoma larva after 4th moult
At – Antenna, An – Antennule, Max I – Maxillipede I, Max II –
Maxillipede II, Max III – Maxillipede III,P1– PereiopodI,P2–
Pereiopod II, P3 – Pereiopod III, P4 – Pereiopod IV, Ab –
Abdomen, CS – Chephalic shield
Fig.6. Percentage of growth rate of phyllosoma larvae
p < 0.0001*** (represents highly significance) done by
regression analysis
Percentage of survival
There was not much mortality up to the first 10days.Forthe
next 10 days, the survival percentage varied between 8.14
and 5.74. After 20 days, the survival percentage started
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declining from 5.39 to 2.67. From 31st day onwards, the
survival percentage further declined and its was 0.01% on
the 42nd day (Fig. 7).
Fig.7. Percentage of survival of phyllosoma larvae
Moulting frequency
The first moult was observed after 9.00 ± 0.57 days of
hatching. The second, third and fourth moultings were
observed on the 19.00 ± 0.57, 28.00 ± 0.57 and 41.33± 0.88
days respectively. There was significant (p < 0.001)
difference between the days taken for each moult up to
fourth (Fig.8).
Fig.8. Moulting frequency of phyllosoma larvae
n=3, p < 0.001** (represents thestatistical significance)done
by ANOVA, followed by Turkey’s multiple comparison tests.
Fecundity
Number of larvae increased significantly (p < 0.001) with
size of the mother lobster (Fig. 9). The spawner of 300gm
size yielded 38,046 ± 26.85 larvae and 400 gm size brooder
released 45,027 ± 20.51 larvae; the brooder of 550 gm size
released 50, 104 ± 57.85 larvae.
Fig.9. Relationship between brooder size and larval
number
n=3, p < 0.001** (represents thestatistical significance)done
by ANOVA, followed by Turkey’s multiple comparison tests
DISCUSSION
When comparing the present result, with Sankolli and
Shenoy (1973) findings, the antennule, antennae, mandible,
first maxilla, second maxilla and second maxilliped had
similar morphological changes. But the following parts
observed by them showed variation with the present work.
The first maxilliped was undeveloped, the third maxilliped
was uniramous, the exopod of first and secondperiopodhad
8 segments and 7 pairs of plumose setae. But in the present
study, the larvae exhibit developed first maxilliped and
biramous third maxilliped. In the presentcase,theexopodof
first and second pereiopod had 7 segments and 6 pairs of
plumose setae each. Again, the present work showedsimilar
morphological changes in the following parts such as eye,
eye stalk, antenna, mandible, firstmaxilla,secondmaxilliped,
and third maxilliped. They observed four apical setae in the
second maxilla, but no such setae were observed in the
present study. Nine pairs of setae were recorded by them in
the first and second pereiopod, but in the present studyonly
7 pairs of setae were observed.
A study conducted by Radhakrishnan and Vijayakumaran
(1993) on phyllosoma larvae of Panulirus homarus showed
un segmented eye stalk during the first moult. The present
study also confirmed their findings. The carapace width
observed by them was 0.72 mm and in the present study it
was 0.68 mm. The early stages of the Panulirus echinatus
moulted eight times and morphological changes of each
appendage were described Abrunhosa et al. (2008). Again a
short span study conducted by Radhakrishnan et al., (2009)
on the survival and growth of phyllosoma larvae of the
tropical spiny lobster, Panulirus homarus maintained with
microalgae Nannochloropsis salina culture treatments and
without microalgae culture, the author reported that the
phyllosoma larvae moulted nine (1- VIII stages) and six
times (I – V stages) in the microalgal and non-algal systems,
respectively. They found the exopod of fourth pereiopod
appeared to become setose in the V stage, the antennule
becomes four segmented in the VI stage, 5th periopod
appeared as small bud in VII stage, 5th pereiopod budding
becomes elongated and formed biramous in stage VIII
respectively. At the end, the total length was 5.25 ± 0.04 and
carapace width was 3.75 ± 0.04 but at the end of the present
study after 4th moult, the total length was 2.89 ± 0.62 mm
and carapace width was 2.01 ± 0.14 mm respectively and
also showed the entire morphological characters of
phyllosoma larvae up to 4th moult.
The spawner size also showed direct relationship with
nauplii or egg production. The same fact has been observed
in P.monodon by (Babu et al., 2001b). About 66% of the
breeders belonged to the size group of61-80mmCLandthis
is the dominant size group in the fishery as evident from the
export of live lobsters. This size group may be contributing
more to the reproduction and recruitment in P. homarus.
This may be due to the deposition of more nutritional
reserve in the body or its increased age.
The fast growth and high survival of early stage phyllosoma
larvae of P. homarus in culture system circulated with N.
salina and continuous enrichment of Artemia by feeding on
N.salina in the micro-algal system was reported by,
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Radhakrishnan et al. (2009). The present study was done
under normal condition. The percentageofsurvivalhadbeen
declining from 31st day onwards. It was also noticed that the
major cause of the larval mortality was the lack of suitable
food for different larval stages (Sarasu and George, 1993;
Abrunhosa, 2008).
ACKNOWLEDGEMENT
The authors are thankful to the University Grants
Commission, Government of India for providing necessary
funds for operating this project and Manonmaniam
Sundaranar University, Tirunelveli,TamilNaduforproviding
Laboratory facilities.
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